Fire Flower selection protocol that removes the shorter DNA fragments from extracted samples

Fire Flower is a user­ friendly spin­ column protocol with a built ­in size depletion ability included in the Fire Monkey kit, and can be used with any other extraction protocol in only 15 minutes.

Fire Flower can be used to size­ select up to 200µl of extracted DNA despite the sample being rich in small DNA fragments. The protocol takes just 15 mins, resulting in an extract which will effectively double most long­-read metrics on LSK109/MinION systems.

Other key features:

  • The Fire Flower size selection protocol is included in the Fire Monkey kit.
  • Fire Flower improves the ratio of high molecular weight versus low molecular weight DNA.
  • Can increase the overall average DNA strand length by 30%.
  • Broad applicability and agnostic of sequencing process used.
  • Can be used with any other extraction protocol.
  • Can be used to size­ select up to 200µl of extracted DNA despite the sample being rich in small DNA fragments.

10-column DNA Size Selection Kit

50-column DNA Size Selection Kit

For Research Use Only. Not intended for diagnostic purposes.

For Research Use Only. Not intended for diagnostic purposes.

FIRE MONKEY & FIRE FLOWER 10 COLUMNS KIT
£195.00 Plus VAT
FIRE MONKEY & FIRE FLOWER 50 COLUMNS KIT
via distributors

Fire Flower

RevoluGen’s Fire Flower protocol by itself can be used independently on any extraction technology and can potentially double the long-read sequencing metrics as shown below

Fire Flower significantly improved the quality of initial samples with 50% short DNA fragments within a short 15 minute protocol...
Key Isolation Data - NanoPlot (LSK109 on MinION, 48hrs)
  Fire Flower Input Fire Flower Output Increase
Average (bp) 17,012 37,163 2.2x
Q-score 10.6 10.7
N50 (bp) 36,919 51,959 1.4x
Total # of reads 215,077 161,295
Total yield (Gb) 3.69 5.99 1.6x
# reads > 50kb 17,460 44,917 2.6x
# reads > 100kb 1,575 3,713 2.4x
# reads > 150kb 123 284 2.3x
Top read 269,583 282,982
% of molecules below and above 30KB1
Fire Flower Input
~ 15mins
Fire Flower Output
  • From an input with only 50% of long DNA extracts, Fire Flower is ablt to deplete small DNA extracts, such that the output contains 80% long DNA extracts.
  • The number of long fragments has more than doubled compared to the input.
1 As per FemtoPulse mesurements
The same result is also seen for samples with an even higher proportion of small DNA fragments
Key Isolation Data - NanoPlot (LSK109 on MinION, 48hrs)
  Fire Flower Input Fire Flower Output Increase
Average (bp) 7,762 20,963 2.7x
Q-score 10.1 10
N50 (bp) 16,901 43,521 2.6x
Total # of reads 1,738,300 191,735
Total yield (Gb) 13.49 4
# reads > 50kb 26,322 23,850 0.9x
# reads > 100kb 1,073 1,110 1.0x
# reads > 150kb 29 45 1.6x
Top read 236,025 213,123
% of molecules below and above 30KB1
Fire Flower Input
~ 15mins
Fire Flower Output
  • Using an even poorer input sample with only 24% long DNA fragments, the Fire Flower is able to deplete short DNA fragments such that the output contains nearly 70% long DNA extracts.
1 As per FemtoPulse mesurements
Fire Flower - validation data
1ml Horse Blood DNA sequenced on R9.4.1/LSK109/MinION for 48hrs - NanoPlot Analysis
Sample Pre-FF Post-FF
Q-score cut-off >7
N50 22,651 40,187
Mean read length (bp) 9,363 11,655
Mean read quality 10.6 9.2
# of reads 1,507,039 1,598,936
Total yield (Total bases) 14,111,188,899 18,635,770,222
Top read (bp)/[Q-score] 301,817/[11.4] 320,028/[9.2]
# of reads > 50,000bp/[~yield in Gb] 43,053/[2.7] 102,105/[6.9]
# of reads > 100,000bp/[~yield in Gb] 2,790/[0.32] 6,108/[0.72]
  • Fire Flower allows the user to achieve yields sufficient to saturate the capacity of the MinION flow cell.
  • Adding only 30mins to the process leads to almost doubling the long read metrics and saturates the flow cell's yield capacity.
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Research & Manufacturing site:

RevoluGen Ltd
Unit 4 Rossington Place
Graphite Way
Hadfield, Derbyshire
SK13 1QG, UK

VAT No. GB 712 2187 67

Head Office:

RevoluGen Ltd
32­-33 High Street
Ascot, Berkshire
SL5 7HG, UK

Registered company No. 03518866
Registered in England and Wales

Follow:
Proud members of:
bia reach ready
bionow
Research & Manufacturing site:

RevoluGen Ltd
Unit 4 Rossington Place
Graphite Way
Hadfield, Derbyshire
SK13 1QG, UK

VAT No. GB 712 2187 67

Head Office:

RevoluGen Ltd
32­-33 High Street
Ascot, Berkshire
SL5 7HG, UK

Registered company No. 03518866
Registered in England and Wales